Human induced pluripotent stem cells for high efficiency genetic engineering
US11866733B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jul 31, 2017 |
| Grant date | Jan 9, 2024 |
| Priority date | — |
| Expiry date | Jul 31, 2037 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2510/00
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Methods are disclosed herein for efficiently generating human induced pluripotent stem cells (iPSC) containing a nucleic acid including a doxycycline promoter operably linked to a nucleic acid encoding Cas9. These methods include transfecting a human somatic cell with a nucleic acid molecule comprising a doxycycline promoter operably linked to a nucleic acid encoding a Cas9, and constitutive promoter operably linked to a tetracycline responsive element and inducing the somatic cell to form an iPSC, thereby producing an iPSC that can undergo CRISPR/Cas9-mediated recombination at a high efficiency. The human iPSC, or a cell differentiated therefrom, is cultured in the presence of doxycycline to induce expression of the Cas9. These cells can then be used to target in any gene of interest by introducing nucleic acids encoding sgRNAs. Induced pluripotent stem cells produced by these methods are also disclosed.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.